Chen Jing, Liu Haimei, Chen Yichuan, Hu Huiyan, Huang Chunxia, Wang Yi, Liang Lijuan, Liu Yunjun
School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, PR China.
School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, PR China; Guangdong Provincial Key Laboratory of Advanced Drug Delivery, Guangdong Provincial Engineering Center of Topical Precise Drug Delivery System, Guangdong Pharmaceutical University, Guangzhou, 510006, PR China.
J Inorg Biochem. 2023 Apr;241:112145. doi: 10.1016/j.jinorgbio.2023.112145. Epub 2023 Jan 23.
Iridium(III) complexes are largely studied as anti-cancer complexes due to their excellent anti-cancer activity. In this article, two new iridium(III) complexes [Ir(piq)(THPIP)]PF (THPIP = 2,4-di-tert-butyl-6-(1H-imidazo[4,5-f][1,10]phenanthrolin-2-yl)phenol, piq = deprotonated 1-phenylisoquinoline) (Ir1) and [Ir(bzq)(THPIP)]PF (bzq = deprotonated benzo[h]quinolone) (Ir2) were synthesized. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays showed that complex Ir1 exhibits moderate activity (IC = 29.9 ± 4.6 μM) and Ir2 shows high cytotoxicity (IC = 9.8 ± 1.8 μM) against BEL-7402 cells. Further studies on the mechanism showed that Ir1 and Ir2 induced apoptosis by changing the mitochondrial membrane potential, Ca release, ROS accumulation, and cell cycle arrest at the S phase. The complexes can effectively inhibit cell colony formation and migration. The expression of B-cell lymphoma-2 (Bcl-2) family proteins, PI3K (phosphatidylinositol 3-kinase), AKT (protein kinase B), mTOR (mammalian target of rapamycin), and p-mTOR was studied by immunoblotting. Complexes Ir1 and Ir2 downregulated the expression of anti-apoptotic protein Bcl-2 and increased the expression of autophagy-related proteins of Beclin-1 and LC3-II. Further experiments showed that the complexes inhibited the production of glutathione (GSH) and increased the amounts of malondialdehyde (MDA). Fluorescence of HMGB1 was significantly increased. We also investigated the effect of the complexes on the expression of genes using RNA-sequence analysis, we further calculated the lowest binding energies between the complexes and proteins using molecular docking. Taken together, the above results indicated that complexes Ir1 and Ir2 induce apoptosis in BEL-7402 cells through a ROS-mediated mitochondrial dysfunction and inhibition of the PI3K/AKT/mTOR signaling pathway.
铱（III）配合物因其出色的抗癌活性而被广泛研究作为抗癌配合物。在本文中，合成了两种新型铱（III）配合物[Ir(piq)(THPIP)]PF（THPIP = 2,4 - 二叔丁基 - 6 -（1H - 咪唑并[4,5 - f][1,10]菲咯啉 - 2 - 基）苯酚，piq = 去质子化的1 - 苯基异喹啉）（Ir1）和[Ir(bzq)(THPIP)]PF（bzq = 去质子化的苯并[h]喹诺酮）（Ir2）。3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐（MTT）实验表明，配合物Ir1对BEL - 7402细胞表现出中等活性（IC = 29.9 ± 4.6 μM），而Ir2显示出高细胞毒性（IC = 9.8 ± 1.8 μM）。对作用机制的进一步研究表明，Ir1和Ir2通过改变线粒体膜电位、钙释放、活性氧积累以及使细胞周期停滞在S期来诱导细胞凋亡。这些配合物能有效抑制细胞集落形成和迁移。通过免疫印迹研究了B细胞淋巴瘤 - 2（Bcl - 2）家族蛋白、磷脂酰肌醇3 - 激酶（PI3K）、蛋白激酶B（AKT）、雷帕霉素靶蛋白（mTOR）和磷酸化mTOR的表达。配合物Ir1和Ir2下调抗凋亡蛋白Bcl - 2的表达，并增加自噬相关蛋白Beclin - 1和LC3 - II的表达。进一步实验表明，这些配合物抑制谷胱甘肽（GSH）的产生并增加丙二醛（MDA）的量。高迁移率族蛋白B1（HMGB1）的荧光显著增加。我们还使用RNA序列分析研究了配合物对基因表达的影响，通过分子对接进一步计算了配合物与蛋白质之间的最低结合能。综上所述，上述结果表明配合物Ir1和Ir2通过活性氧介导的线粒体功能障碍和抑制PI3K/AKT/mTOR信号通路诱导BEL - 7402细胞凋亡。
